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Acknowledgements
This method was adapted from the Cell Signaling Consortium protocol for transfecting 293T cells in suspension (kindly provided by Iain Fraser; California Institute of Technology) and the manufacturer's protocol for Lipofectamine 2000. The BAC recombination protocol and shuttle vector for creating the Nanog-GFP construct were kindly provided by Nathanial Heintz (Rockefeller University). P.J.P. thanks A. Mills and her lab at Cold Spring Harbor Laboratory (CSHL) for help with ES cell culture. P.J.P. is supported by the CSHL President's Council Fund for CSHL fellows.
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Schaniel, C., Li, F., Schafer, X. et al. Delivery of short hairpin RNAs—triggers of gene silencing—into mouse embryonic stem cells. Nat Methods 3, 397–400 (2006). https://doi.org/10.1038/nmeth0506-397
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DOI: https://doi.org/10.1038/nmeth0506-397
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